A kind of method utilizing bioreactor to prepare porcine circovirus type 2 inactivated vaccine

Abstract

The invention relates to a method for preparing porcine circovirus type 2 inactivated vaccine by utilizing a bioreactor, wherein the bioreactor is a riptide type bioreactor; a circulatory system is composed of a perfusion bag and a riptide bag; a cell culture bag is built in the perfusion bag; a paper carrier is in the cell culture bag; and the method comprises the following steps of: culturing PK-15 cells; inoculating porcine circovirus type 2; culturing the porcine circovirus type 2, measuring sugar consumption by taking a virus maintenance solution, and obtaining a virus solution when the sugar consumption is decreased; and preparing the porcine circovirus type 2 inactivated vaccine. The method for preparing the porcine circovirus type 2 inactivated vaccine by utilizing the riptide type bioreactor disclosed by the invention is small in production site, low in labour intensity, relatively lower in production cost, simple in operation, low in inter-batch difference, and high and steady in product quality; full-automatic micro-computer control can be achieved; and the prepared porcine circovirus type 2 inactivated vaccine is good in safety, high in immune efficacy and low in side effect and has the better immune protective effect on virulent attack of porcine circovirus type 2.

Description

technical field [0001] The invention relates to a preparation method of a vaccine, in particular to a method for preparing porcine circovirus type 2 inactivated vaccine by utilizing a bioreactor. Background technique [0002] Porcine circovirus (porcine circovirus, PCV) belongs to the genus Circovirus in the family Circoviridae in taxonomy, and is one of the smallest known animal viruses. Virus particles have a diameter of 14-17nm, a 20-sided symmetrical structure, no envelope, and contain a covalently closed single-strand circular negative-strand DNA. The genome size is about 1.76kb. PCV is quite resistant to external physical and chemical factors, and can survive for a period of time even in an acidic environment of PH3 and a high temperature environment of 72°C. It is not inactivated by the action of chloroform, and has no hemagglutination activity. PCV is known to have two serotypes, PCV1 and PCV2. PCV1 is a non-pathogenic virus. PCV2 is a pathogenic virus. [0003] ...

AI Technical Summary

Problems solved by technology

However, the proliferation of spinner bottle cultured cells is relatively slow

Moreover, the cultivation conditions of cells and viruses in the production process, such as pH, dissolved oxygen, sugar consumption, etc., are difficult to monitor and timely supply, and cannot provide the best cultivation conditions, resulting in a low degree of automation of the method, high labor intensity, and because of bottle rotation The uncontrollability of the cell culture environment leads to insufficient quality stability of the produced products and large batch-to-batch differences

In addition, because the virus is a DNA virus, the mutation rate of the virus is low, and at the same time, only PCV2 is the only cause of disease in the world.

However, the biological specificity of PCV2 is quite special. Its proliferation titer on cells is very low, and it does not cause cytopathic changes. Therefore, the low titer of toxin production is also a more important problem, and it is not compatible with the current large-scale animal vaccine production. not compatible

In addition, if you want to expand the production scale, you can only increase the number of bottles, which results in greater demand for workshop scale, equipment access, personnel, etc., and greater labor.

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